D-homoandrosta-17-yl carbamate derivatives as selective glucocorticoid receptor ligands

ABSTRACT

The present invention is directed to D-homoandrosta-17-yl-carbamate derivatives as selective glucocorticoid receptor ligands useful for treating a variety of autoimmune and inflammatory diseases or conditions. Pharmaceutical compositions and methods of use are also included.

BACKGROUND OF THE INVENTION

Intracellular receptors (IR's) are a class of structurally relatedproteins involved in the regulation of gene expression. The steroidhormone receptors are a subset of this superfamily whose natural ligandsare typically comprised of endogenous steroids such as estradiol,progesterone, and cortisol. Man-made ligands to these receptors play animportant role in human health and, of these receptors, theglucocorticoid receptor has an essential role in regulating humanphysiology and immune response. Steroids that interact with theglucocorticoid receptor have been shown to be potent anti-inflammatoryagents. The present invention is directed to a novel class of compoundsthat are selective glucocorticoid receptor modulators that have potentanti-inflammatory and immunosuppressive activity and possess advantagesover steroidal glucocorticoid ligands with respect to side effects,efficacy, toxicity and/or metabolism.

SUMMARY OF THE INVENTION

The present invention is directed to D-homoandrosta-17-yl-carbamatederivatives as selective glucocorticoid receptor ligands useful fortreating a variety of autoimmune and inflammatory diseases orconditions. Pharmaceutical compositions and methods of use are alsoincluded.

DETAILED DESCRIPTION OF THE INVENTION

The invention encompasses a genus of compounds of Formula I

or a pharmaceutically acceptable salt thereof, whereinR¹ and R² are independently selected from the group consisting of:R^(a), —C(O)—R^(a), and —C(O)—N(R^(a))R^(a);R³ and R⁴ are independently selected from the group consisting of:hydrogen, C₁₋₁₀alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₃₋₁₀cycloalkyl, aryl,hetreoaryl, heterocyclyl, and C₁₋₁₀fluoroalkyl; andeach Ra is independently selected from the group consisting of:hydrogen, C₁₋₁₀alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₃₋₆cycloalkyl, aryl,hetreoaryl, heterocyclyl, and C₁₋₁₀fluoroalkyl.

Within the genus, the invention encompasses a sub-genus of compounds ofFormula I wherein R² is selected from hydrogen and CH₃—C(O)—.

Also within the genus, the invention encompasses a sub-genus ofcompounds of Formula I wherein R¹ is selected from hydrogen andCF₃—(CO)—.

Also within the genus, the invention encompasses a sub-genus ofcompounds of Formula I wherein R³ is hydrogen. Within this sub-genus,the invention encompasses a class of compounds of Formula I wherein R⁴is selected from C₁₋₁₀alkyl and C₃₋₁₀cycloalkyl.

Also within the genus, the invention encompasses a sub-genus ofcompounds of Formula I wherein: R¹ is selected from hydrogen orCF₃—(CO)—; R² is selected from hydrogen and CH₃—C(O)—; R³ is hydrogen;and R⁴ is selected from C₁₋₁₀alkyl and C₃₋₁₀cycloalkyl.

Within this sub-genus, the invention encompasses a class of compounds ofFormula I wherein R⁴ is C₁₋₆alkyl.

Also within this sub-genus, the invention encompasses a class ofcompounds of Formula I wherein R⁴ is selected from: cyclopentyl,cyclohexyl and adamantanyl.

Also within this sub-genus, the invention encompasses a class ofcompounds of Formula I wherein R¹ and R² are hydrogen.

Illustrating the invention are the following compounds:

-   (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate,-   (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(2-pentyl)carbamate,-   (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(2-methyl-pentyl)carbamate,-   (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-7-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(3-methyl-2-propyl)carbamate,-   (11β,16β,17α)-9-fluoro-11-trifluoroacetoxy-16-methyl-3,17a-dioxo-17-[(acetyloxy)methyl]-D-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate,-   (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-[(acetyloxy)methyl]-D-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate,-   (11β,16β,17α)-9-fluoro-11-trifluoroacetoxy    16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate,-   (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(cyclopentyl)carbamate,-   (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(cyclohexyl)carbamate    or a pharmaceutically acceptable salt of any of the above.

Another embodiment of the invention encompasses a pharmaceuticalcomposition comprising a compound of Formula I in combination with apharmaceutically acceptable carrier.

Another embodiment of the invention encompasses a method for treating aglucocorticoid receptor mediated disease or condition in a mammalianpatient in need of such treatment comprising administering the patient acompound of Formula I in an amount that is effective for treating theglucocorticoid receptor mediated disease or condition.

Within this embodiment is encompassed the above method wherein theglucocorticoid receptor mediated disease or condition is selected fromthe group consisting of: tissue rejection, leukemias, lymphomas,Cushing's syndrome, acute adrenal insufficiency, congenital adrenalhyperplasia, rheumatic fever, polyarteritis nodosa, granulomatouspolyarteritis, inhibition of myeloid cell lines, immuneproliferation/apoptosis, ETA axis suppression and regulation,hypercortisolemia, stroke and spinal cord injury, hypercalcemia,hypergylcemia, acute adrenal insufficiency, chronic primary adrenalinsufficiency, secondary adrenal insufficiency, congenital adrenalhyperplasia, cerebral edema, thrombocytopenia, Little's syndrome,obesity, metabolic syndrome, inflammatory bowel disease, systemic lupuserythematosus, polyartitis nodosa, Wegener's granulomatosis, giant cellarteritis, rheumatoid arthritis, juvenile rheumatoid arthritis, uveitis,hay fever, allergic rhinitis, urticaria, angioneurotic edema, chronicobstructive pulmonary disease, asthma, tendonitis, bursitis, Crohn'sdisease, ulcerative colitis, autoimmune chronic active hepatitis, organtransplantation, hepatitis, cirrhosis, inflammatory scalp alopecia,panniculitis, psoriasis, discoid lupus erythematosus, inflamed cysts,atopic dermatitis, pyoderma gangrenosum, pemphigus vulgaris, buflouspernphigoid, systemic lupus erythematosus, dermatomyositis, herpesgestationis, eosinophilic fasciitis, relapsing polychondritis,inflammatory vasculitis, sarcoidosis, Sweet's disease, type I reactiveleprosy, capillary hemangiomas, contact dermatitis, atopic dermatitis,lichen planus, exfoliative dermatitus, erythema nodosum, acne,hirsutism, toxic epidermal necrolysis, erythema multiform, cutaneousT-cell lymphoma, Human Immunodeficiency Virus (HIV), cell apoptosis,cancer, Kaposi's sarcoma, retinitis pigmentosa, cognitive performance,memory and learning enhancement, depression, addiction, mood disorders,chronic fatigue syndrome, schizophrenia, sleep disorders, and anxiety.

Another embodiment of the invention encompasses a method of selectivelymodulating the activation, repression, agonism and antagonism effects ofthe glucocorticoid receptor in a mammal comprising administering to themammal a compound of Formula I in an amount that is effective tomodulate the glucocorticoid receptor.

Exemplifying the invention are the compounds of the Examples disclosedhereunder.

DEFINITIONS

The invention is described using the following definitions unlessotherwise indicated.

“Alkyl”, as well as other groups having the prefix “alk”, such asalkoxy, alkanoyl, means carbon chains which may be linear or branched orcombinations thereof. Examples of alkyl groups include methyl, ethyl,propyl, isopropyl, butyl, sec- and tert-butyl, pentyl, hexyl, heptyl,octyl, nonyl, and the like.

“Fluoroalkyl” means alkyl as defined above wherein one or more of thehydrogen atoms are replaced with a fluoro atom, up to the maximum numberof substitutable positions.

“Alkenyl” means carbon chains which contain at least one carbon-carbondouble bond, and which may be linear or branched or combinationsthereof. Examples of alkenyl include vinyl, allyl, isopropenyl,pentenyl, hexenyl, heptenyl, 1-propenyl, 2-butenyl, 2-methyl-2-butenyl,and the like.

“Alkynyl” means carbon chains which contain at least one carbon-carbontriple bond, and which may be linear or branched or combinationsthereof. Examples of alkynyl include ethynyl, propargyl,3-methyl-1-pentynyl, 2-heptynyl and the like.

“Cycloalkyl” means mono-, bi- or tri-cyclic saturated carbocyclic ringshaving the indicated number of carbon atoms. The term also includesmonocyclic rings fused to an aryl group in which the point of attachmentis on the non-aromatic portion. Examples of cycloalkyl includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,tetrahydronaphthyl, decahydronaphthyl, indanyl, adamantanyl and thelike.

“Aryl” means mono- or bicyclic aromatic rings containing only carbonatoms. The term also includes aryl group fused to a monocycliccycloalkyl or monocyclic heterocyclyl group in which the point ofattachment is on the aromatic portion. Examples of aryl include phenyl,naphthyl, indanyl, indenyl, tetrahydronaphthyl, 2,3-dihydrobenzofuranyl,dihydrobenzopyranyl, 1,4-benzodioxanyl, and the like.

“Heteroaryl” means a mono- or bicyclic aromatic ring containing at leastone heteroatom selected from N, O and S, with each ring containing 5 to6 atoms. Examples of heteroaryl include pyrrolyl, isoxazolyl,isothiazolyl, pyrazolyl, pyridyl, oxazolyl, oxadiazolyl, thiadiazolyl,thiazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, triazinyl,thienyl, pyrimidyl, pyridazinyl, pyrazinyl, benzoxazolyl,benzothiazolyl, benzimidazolyl, benzofuranyl, benzothiophenyl,furo(2,3-b)pyridyl, quinolyl, indolyl, isoquinolyl, and the like.

“Heterocyclyl” means mono- or bicyclic saturated rings containing atleast one heteroatom selected from N, S and O, each of said ring havingfrom 3 to 10 atoms in which the point of attachment may be carbon ornitrogen. The term also includes monocyclic heterocycle fused to an arylor heteroaryl group in which the point of attachment is on thenon-aromatic portion. Examples of “heterocyclyl” include pyrrolidinyl,piperidinyl, piperazinyl, imidazolidinyl, 2,3-dihydrofuro(2,3-b)pyridyl,benzoxazinyl, tetrahydrohydroquinolinyl, tetrahydroisoquinolinyl,dihydroindolyl, and the like. The term also includes partiallyunsaturated monocyclic rings that are not aromatic, such as 2- or4-pyridones attached through the nitrogen orN-substituted-(1H,3H)-pyrimidine-2,4-diones (N-substituted uracils).

Abbreviations

The following abbreviations have the indicated meanings:

AIBN=2,2′-azobisisobutyronitrile

B.P.=benzoyl peroxide

Bn=benzyl

CCl₄=carbon tetrachloride

D=—O(CH₂)₃O—

DAST=diethylamine sulfur trifluoride

DCC=dicyclohexyl carbodiimide

DCI=1-(3-dimethylaminopropyl)-3-ethyl carbodiimide

DEAD=diethyl azodicarboxylate

DIBAL=diisobutyl aluminum hydride

DME=ethylene glycol dimethylether

DMAP=4-(dimethylamino)pyridine

DMF=N,N-dimethylformamide

DMSO=dimethyl sulfoxide

Et₃N=triethylamine

LDA=lithium diisopropylamide

m-CPBA=metachloroperbenzoic acid

NBS=N-bromosuccinimide

NSAID=non-steroidal anti-inflammatory drug

PCC=pyridinium chlorochromate

PDC=pyridinium dichromate

Ph=phenyl

1,2-Ph=1,2-benzenediyl

Pyr=pyridinediyl

Qn=7-chloroquinolin-2-yl

R^(S)=—CH₂SCH₂CH₂Ph

r.t.=room temperature

rac.=racemic

THF=tetrahydrofuran

THP=tetrahydropyran-2-yl

Alkyl Group Abbreviations

Me=methyl

Et=ethyl

n-Pr=normal propyl

i-Pr=isopropyl

n-Bu=normal butyl

i-Bu=isobutyl

s-Bu=secondary butyl

t-Bu=tertiary butyl

c-Pr=cyclopropyl

c-Bu=cyclobutyl

c-Pen=cyclopentyl

c-Hex=cyclohexyl

Optical Isomers—Diastereomers—Geometric Isomers—Tautomers

Compounds of Formula I contain one or more asymmetric centers and canthus occur as racemates and racemic mixtures, single enantiomers,diastereomeric mixtures and individual diastereomers. The presentinvention is meant to comprehend all such isomeric forms of thecompounds of Formula I.

Some of the compounds described herein contain olefinic double bonds,and unless specified otherwise, are meant to include both E and Zgeometric isomers.

Some of the compounds described herein may exist with different pointsof attachment of hydrogen, referred to as tautomers. Such an example maybe a ketone and its enol form known as keto-enol tautomers. Theindividual tautomers as well as mixture thereof are encompassed withcompounds of Formula I.

Compounds of the Formula I may be separated into diastereoisomeric pairsof enantiomers by, for example, fractional crystallization from asuitable solvent, for example MeOH or EtOAc or a mixture thereof. Thepair of enantiomers thus obtained may be separated into individualstereoisomers by conventional means, for example by the use of anoptically active amine as a resolving agent or on a chiral HPLC column.

Alternatively, any enantiomer of a compound of the general Formula I orIa may be obtained by stereospecific synthesis using optically purestarting materials or reagents of known configuration.

Salts

The term “pharmaceutically acceptable salts” refers to salts preparedfrom pharmaceutically acceptable non-toxic bases or acids includinginorganic or organic bases and inorganic or organic acids. Salts derivedfrom inorganic bases include aluminum, ammonium, calcium, copper,ferric, ferrous, lithium, magnesium, manganic salts, manganous,potassium, sodium, zinc, and the like. Particularly preferred are theammonium, calcium, magnesium, potassium, and sodium salts. Salts derivedfrom pharmaceutically acceptable organic non-toxic bases include saltsof primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines, and basic ionexchange resins, such as arginine, betaine, caffeine, choline,N,N-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol,2-dimethylaminoethanol, ethanolamine, ethylenediamine,N-ethyl-morpholine, N-ethylpiperidine, glucamine, glucosamine,histidine, hydrabamine, isopropylamine, lysine, methylglucamine,morpholine, piperazine, piperidine, polyamine resins, procaine, purines,theobromine, triethylamine, trimethylamine, tripropylamine,tromethamine, and the like.

When the compound of the present invention is basic, salts may beprepared from pharmaceutically acceptable non-toxic acids, includinginorganic and organic acids. Such acids include acetic, benzenesulfonic,benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic,glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic,mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic,phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid, andthe like. Particularly preferred are citric, hydrobromic, hydrochloric,maleic, phosphoric, sulfuric, and tartaric acids.

Dose Ranges

It will be understood that, as used herein, references to the compoundsof Formula I are meant to also include the pharmaceutically acceptablesalts.

The magnitude of prophylactic or therapeutic dose of a compound ofFormula I will, of course, vary with the nature and the severity of thecondition to be treated and with the particular compound of Formula Iand its route of administration. It will also vary according to avariety of factors including the age, weight, general health, sex, diet,time of administration, rate of excretion, drug combination and responseof the individual patient. In general, the daily dose from about 0.001mg to about 100 mg per kg body weight of a mammal, preferably 0.01 mg toabout 10 mg per kg. On the other hand, it may be necessary to usedosages outside these limits in some cases.

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. For example, aformulation intended for oral administration to humans may contain fromabout 0.5 mg to about 5 g of active agent compounded with an appropriateand convenient amount of carrier material which may vary from about 5 toabout 95 percent of the total composition. Dosage unit forms willgenerally contain from about 1 mg to about 2 g of an active ingredient,typically 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg,800 mg, or 1000 mg.

Pharmaceutical Compositions

For the treatment of glucocorticoid receptor mediated diseases thecompound of Formula I may be administered orally, topically,parenterally, by inhalation spray or rectally in dosage unitformulations containing conventional non-toxic pharmaceuticallyacceptable carriers, adjuvants and vehicles. The term parenteral as usedherein includes subcutaneous, intravenous, intramuscular, intrasternalinjection or infusion techniques. In addition to the treatment ofwarm-blooded animals such as mice, rats, horses, cattle, sheep, dogs,cats, etc., the compound of the invention is effective in the treatmentof humans.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, solutions, aqueous or oily suspensions, dispersible powders orgranules, emulsions, hard or soft capsules, syrups or elixirs.Compositions intended for oral use may be prepared according to anymethod known to the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more agentsselected from the group consisting of sweetening agents, flavouringagents, colouring agents and preserving agents in order to providepharmaceutically elegant and palatable preparations. Tablets contain theactive ingredient in admixture with non-toxic pharmaceuticallyacceptable excipients which are suitable for the manufacture of tablets.These excipients may be for example, inert diluents, such as calciumcarbonate, sodium carbonate, lactose, calcium phosphate or sodiumphosphate; granulating and disintegrating agents, for example, cornstarch, or alginic acid; binding agents, for example starch, gelatin oracacia, and lubricating agents, for example, magnesium stearate, stearicacid or talc. The tablets may be uncoated or they may be coated by knowntechniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate may be employed. They may also becoated by the technique described in the U.S. Pat. Nos. 4,256,108;4,166,452; and 4,265,874 to form osmotic therapeutic tablets for controlrelease.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredients is mixed withwater-miscible solvents such as propylene glycol, PEGs and ethanol, oran oil medium, for example peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active material in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more colouringagents, one or more flavouring agents, and one or more sweeteningagents, such as sucrose, saccharin or aspartame.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavouring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavouring and colouringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof an oil-in-water emulsion. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavouring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavouring and colouringagents. The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. Cosolvents suchas ethanol, propylene glycol or polyethylene glycols may also be used.In addition, sterile, fixed oils are conventionally employed as asolvent or suspending medium. For this purpose any bland fixed oil maybe employed including synthetic mono- or diglycerides. In addition,fatty acids such as oleic acid find use in the preparation ofinjectables.

The compounds of Formula I may also be administered in the form ofsuppositories for rectal administration of the drug. These compositionscan be prepared by mixing the drug with a suitable non-irritatingexcipient which is solid at ambient temperatures but liquid at therectal temperature and will therefore melt in the rectum to release thedrug. Such materials are cocoa butter and polyethylene glycols.

For topical use, creams, ointments, gels, solutions or suspensions,etc., containing a compound of Formula I are employed. (For purposes ofthis application, topical application shall include mouth washes andgargles.) Topical formulations may generally be comprised of apharmaceutical carrier, cosolvent, emulsifier, penetration enhancer,preservative system, and emollient.

Utilities

The ability of the compounds of Formula I to selectively modulateglucocorticoid receptors makes them useful for treating, preventing orreversing the progression of a variety of inflammatory and autoimmunediseases and conditions. Thus, the compounds of the present inventionare useful to treat, prevent or ameliorate the following diseases orconditions: inflammation, tissue rejection, auto-immunity, variousmalignancies, such as leukemias and lymphomas, Cushing's syndrome, acuteadrenal insufficiency, congenital adrenal hyperplasia, rheumatic fever,polyarteritis nodosa, granulomatous polyarteritis, inhibition of myeloidcell lines, immune proliferation/apoptosis, HPA axis suppression andregulation, hypercortisolemia, stroke and spinal cord injury,hypercalcemia, hypergylcemia, acute adrenal insufficiency, chronicprimary adrenal insufficiency, secondary adrenal insufficiency,congenital adrenal hyperplasia, cerebral edema, thrombocytopenia,Little's syndrome, obesity and metabolic syndrome.

The compounds of the present invention are also useful for treating,preventing or reversing the progression of disease states involvingsystemic inflammation such as inflammatory bowel disease, systemic lupuserythematosus, polyartitis nodosa, Wegener's granulomatosis, giant cellarteritis, rheumatoid arthritis, juvenile rheumatoid arthritis, uveitis,hay fever, allergic rhinitis, urticaria, angioneurotic edema, chronicobstructive pulmonary disease, asthma, tendonitis, bursitis, Crohn'sdisease, ulcerative colitis, autoimmune chronic active hepatitis, organtransplantation, hepatitis, and cirrhosis. The compounds of the presentinvention are useful for treating, preventing or reversing theprogression of a variety of topical diseases such as inflammatory scalpalopecia, panniculitis, psoriasis, discoid lupus erythematosus, inflamedcysts, atopic dermatitis, pyoderma gangrenosum, pemphigus vulgaris,buflous pernphigoid, systemic lupus erythematosus, dermatomyositis,herpes gestation is, eosinophilic fasciitis, relapsing polychondritis,inflammatory vasculitis, sarcoidosis, Sweet's disease, type I reactiveleprosy, capillary hemangiomas, contact dermatitis, atopic dermatitis,lichen planus, exfoliative dermatitus, erythema nodosum, acne,hirsutism, toxic epidermal necrolysis, erythema multiform, cutaneousT-cell lymphoma.

The compounds of the present invention are also useful in treating,preventing or reversing the progression of disease states associatedwith Human Immunodeficiency Virus (HIV), cell apoptosis, and cancerincluding, but not limited to, Kaposi's sarcoma, immune systemactivation and modulation, desensitization of inflammatory responses,IIL-I expression, natural killer cell development, lymphocytic leukemia,and treatment of retinitis pigmentosa. Cognitive and behavioralprocesses are also susceptible to glucocorticoid therapy whereantagonists would potentially be useful in the treatment of processessuch as cognitive performance, memory and learning enhancement,depression, addiction, mood disorders, chronic fatigue syndrome,schizophrenia, stroke, sleep disorders, and anxiety.

Preferably, the compounds of the invention are useful for treating thediseases or conditions set for the below.

1. Allergic States

Control of severe or incapacitating allergic conditions not responsiveto adequate trials of conventional treatment; seasonal or perennialallergic rhinitis; bronchial asthma; contact dermatitis; atopicdermatitis; serum sickness; and drug hypersensitivity reactions.

2. Rheumatic Disorders

As adjunctive therapy for short-term administration during an acuteepisode or exacerbation of: psoriatic arthritis; rheumatoid arthritisincluding juvenile rheumatoid arthritis (selected cases may requirelow-dose maintenance therapy); ankylosing spondylitis; acute andsubacute bursitis; acute nonspecific tenosynovitis; acute goutyarthritis; post-traumatic osteoarthritis; synovitis of osteoarthritis;and epicondylitis

3. Dermatologic Diseases

Pemphigus; bullous dermatitis herpetiformis; severe erythema multiforme(Stevens-Johnson syndrome); exfoliative dermatitis; mycosis fungoïdes;severe psoriasis; and severe seborrheic dermatitis.

4. Ophthalmic Diseases

Severe acute and chronic allergic and inflammatory processes involvingthe eye and its adnexa such as: allergic conjunctivitis; keratitis;allergic corneal marginal ulcers; herpes zoster ophthalmicus; iritis andiridocyclitis; chorioretinitis; anterior segment inflammation; diffuseposterior uveitis and choroiditis; optic neuritis; and sympatheticophthalmia

5. Endocrine Disorders

Primary or secondary adrenocortical insufficiency; congenital adrenalhyperplasia; nonsuppurative thyroiditis; and hypercalcemia associatedwith cancer.

6. Respiratory Diseases

Symptomatic sarcoidosis; Löffler's syndrome not manageable by othermeans; berylliosis; fulminating or disseminated pulmonary tuberculosiswhen concurrently accompanied by appropriate antituberculouschemotherapy; and aspiration pneumonitis.

7. Hematologic Disorders

Idiopathic thrombocytopenic purpura in adults; secondarythrombocytopenia in adults; acquired (autoimmune) hemolytic anemia;erythroblastopenia (RBC anemia); and congenital (erythroid) hypoplasticanemia.

8. Neoplastic Diseases

For palliative management of: leukemias and lymphomas in adults; andacute leukemia of childhood.

9. Edematous States

To induce a diuresis or remission of proteinuria in the nephroticsyndrome without uremia, of the idiopathic type or that due to lupuserythematosus. Compounds of Formula I may be used to treat patients withcerebral edema from various causes. It may be used also in thepreoperative preparation of patients with increased intracranialpressure secondary to brain tumors, and also for palliation of patientswith inoperable or recurrent brain neoplasms, and in the management ofcerebral edema associated with neurosurgery. Some patients with cerebraledema due to head injury or pseudotumor cerebri also may benefit fromtherapy with compounds of Formula I.

10. Gastrointestinal Diseases

During a critical period of the disease in: ulcerative colitis andregional enteritis.

11. Miscellaneous

Tuberculous meningitis with subarachnoid block or impending block whenconcurrently accompanied by appropriate antituberculous chemotherapy;Trichinosis with neurologic or myocardial involvement; During anexacerbation or as maintenance therapy in selected cases of: Systemiclupus erythematosus and acute rheumatic carditis; in combination withondansetron for the management of nausea and vomiting associated withcisplatin and non-cisplatin emetogenic chemotherapy.

Combination Therapy

The invention also encompasses a method for treating a glucocorticoidreceptor mediated disease comprising concomitantly administering to apatient in need of such treatment a compound of Formula I and one oradditional more agents. For treating or preventing asthma or chronicobstructive pulmonary disease, the compounds of Formula I may becombined with one or more agents selected from the group consisting of:θ-agonists (e.g., salmeterol), theophylline, anticholinergics (e.g.,atropine and ipratropium bromide), cromolyn, nedocromil and leukotrienemodifiers (e.g., montelukast). For treating or preventing inflammation,the compounds of Formula I may be combined with one or the following: asalicylate, including acetylsalicylic acid, a non-steroidalantiinflammatory drug, including indomethacin, sulindac, mefenamic,meclofenamic, tolfenamic, tolmetin, ketorolac, dicofenac, ibuprofen,naproxen, fenoprofen, ketoprofen, flurbiprofin and oxaprozin, a TNFinhibitor, including etanercept and infliximab, an IL-1 receptorantagonist, a cytotoxic or immunosuppressive drug, includingmethotrexate, leflunomide, azathioprine and cyclosporine, a goldcompound, hydroxychloroquine or sulfasalazine, penicillamine,darbufelone, and a ρ38 kinase inhibitor. The compound of Formula I mayalso be used in combination with bisphonates such as alendronate totreat a glucocorticoid mediated disease and simultaneously inhibitosteoclast-mediated bone resorption.

Methods of Synthesis and Examples

The invention will now be illustrated by the following non-limitingexamples in which, unless stated otherwise:

(i) all operations were carried out at room or ambient temperature, thatis, at a temperature in the range 18-25° C.,

(ii) evaporation of solvent was carried out using a rotary evaporatorunder reduced pressure (600-4000 pascals: 4.5-30 mm. Hg) with a bathtemperature of up to 60° C.,

(iii) the course of reactions was followed by thin layer chromatography(TLC) and reaction times are given for illustration only;

(iv) melting points are uncorrected and ‘d’ indicates decomposition; themelting points given are those obtained for the materials prepared asdescribed; polymorphism may result in isolation of materials withdifferent melting points in some preparations;

(v) the structure and purity of all final products were assured by atleast one of the following techniques: TLC, mass spectrometry, nuclearmagnetic resonance (NMR) spectrometry or microanalytical data;

(vi) yields are given for illustration only;

(vii) when given, NMR data is in the form of delta (δ) values for majordiagnostic protons, given in parts per million (ppm) relative totetramethylsilane (TMS) as internal standard, determined at 500 MHz or600 MHz using the indicated solvent; conventional abbreviations used forsignal shape are: s. singlet; d. doublet; t. triplet; m. multiplet; br.broad; etc.: in addition “Ar” signifies an aromatic signal;

(viii) chemical symbols have their usual meanings; the followingabbreviations have also been used v (volume), w (weight), b.p. (boilingpoint), m.p. (melting point), L (litre(s)), mL (millilitres), g(gram(s)), mg (milligrams(s)), mol (moles), mmol (millimoles), eq(equivalent(s)).

Example 1 Synthesis of(11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-d-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate

(11β,16β)-9-fluoro-17,21-dihydroxy-16-methyl-3,20-dioxo-11-trifluoroacetoxy-pregna-1,4-diene(1-3)

Betamethasone (1-1, 96 g, 245 mmol) and DMAP (6.0 g, 49 mmol) weredissolved in 2.8 L of dry THF with stirring. The solution was placed inan ice bath and cooled to 10-15° C. To the cooled solution was addedtrifluoroacetic anhydride (129 g, 614 mmol) via an addition funnel overa 20 minute period. The ice bath was allowed to expire and the solutionwas allowed to stir at room temperature for 15 hours. To the solutionwas added 675 mL of water, and the resulting mixture was allowed to stirfor 15 hours. The mixture was reduced to one-quarter the original volumein vacuo and the residue was diluted with 2 L of water. The precipitatewas isolated by filtration and dried for 15 hrs at reduced pressure toproduce 122.5 g of 1-3 as a pale white solid.

MS (ESI): m/z=489.2 (MH⁺).

(11β,16β)-9-fluoro-17-hydroxy-16-methyl-3,20-dioxo-11-trifluoroacetoxy-21-acetoxy-pregna-1,4-diene(1-4)

To a solution of 1-3 (122.5 g) in THF (1 L) at RT was added DMAP (6.0 g,49 mmol) and acetic anhydride (50.2 g, 491 mmol) and the mixture allowedto stir for 15 hours. The solvent was evaporated, and the residuediluted with 1 L of water. The precipitate was isolated by filtrationand dried for 15 hrs in a warm (35° C.) vacuum oven to produce 123.4 g(94.7%) 1-4 as an off-white solid.

MS (ESI): m/z=531.2 (MH⁺).

(11β,16β,17α)-9-fluoro-17-hydroxy-11-trifluoroacetoxy-16-methyl-3,17a-dioxo-17-[(acetyloxy)methyl]-D-homoandrosta-1,4-diene(1-5)

A suspension of 1-4 (42.0 g, 79.168 mmol) in 630 mL anhydrousacetonitrile was cooled to −5° C. and SnCl₄ (41.24 g, 158.335 mmol) wasadded dropwise. The resulting homogenous solution was warmed to ambienttemperature stirred for 72 h during which time a white solidprecipitated. The solid was filtered and dried overnight at 35° C. togive 34.10 g of 1-5. The filtrate was diluted with ethyl acetate (2 L)and diluted with a mixture of sodium bicarbonate (6.651 g, 79.168 mmol)in 300 mL water. The combined organic layers were washed with water,saturated brine solution, dried over sodium sulfate, filtered andconcentrated in vacuo to give an additional 3 g of 1-5 as beige foamysolid.

MS (ESI): m/z=531.3 (MH+).

(11β,16β,17α)-9-fluoro-11-trifluoroacetoxy-16-methyl-3,17a-dioxo-17-[(acetyloxy)methyl]-D-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate(1-6)

A solution of 1-5 (0.815 g, 1.536 mmol) in 2.2 mL anhydrousN,N-dimethylformamide was purged with a stream of nitrogen for 10minutes, then 1-adamantyl isocyanate (0.545 g, 3.072 mmol) and CuCl(0.152 g, 1.536 mmol) were added. The resulting green solution wasstirred at ambient temperature for 1 h. The reaction mixture was pouredinto n5 mL water and extracted with ethyl acetate (2×50 mL). Thecombined organic layers were washed with 10 mL water, 10 mL saturatedbrine solution, dried over magnesium sulfate, filtered, and concentratedin vacuo to give 1.18 g of an orange viscous oil. The oil was dissolvedin with toluene (15 mL) and concentrated in vacuo to give a pale greensolid (1.00 g). The residue was subjected to flash chromatographythrough a column of 40 g of silica gel, eluting with a gradient of 0%B/100% A to 20% B/80% A over 55 min (A=50% dichloromethane/50% hexane,B=ethyl acetate) to provide 0.646 g (59%) of 1-6 as a beige solid.

MS (ESI): m/z=708.3 (MH+).

(11β,16β,17α)-9-fluoro-11-trifluoroacetoxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4dien-17-yl-(1-adamantyl)carbamate(1-7)

Solid 1-6 (610 g, 0.862 mmol) was dissolved in 30 mL of a premixedsolution of methanol/chloroform 6N HCl (10:2:1) and the resultingmixture stirred at ambient temperature for 15 hours. An additional 1 mLof 6N HCl was added and the mixture stirred for a further 20 hours. Themixture was concentrated in vacuo to give 0.544 g of 1-7 as a beigesolid.

MS (ESI): m/z=666.3 (MH+).

(11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate(1-8)

To a solution of 1-7 (0.492 g, 0.739 mmol) in 36 mL methanol was addedtriethylamine (0.206 mL, 1.478 mmol) and the resulting colorlesssolution was stirred at ambient temperature for 30 min then concentratedin vacuo to give a beige foamy solid. The solid was dissolved 7 mL hotdichloromethane, loaded onto a 40 g silica gel cartridge, and elutedwith 0% B/100% A for 10 min, then with a gradient of 0% B/100% A to 50%B/50% A over 50 min to yield 0.330 g 1-8 as a white solid (78%).

MS (ESI): m/z=570.3 (MH+).

(11β,16β,17β)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-[(acetyloxy)methyl]-D-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate(1-9)

To a solution of 1-6 (0.100 g, 0.141 mmol) in 7 mL methanol was addedtriethylamine (0.039 mL, 0.283 mmol) and the resulting colorlesssolution was stirred at ambient temperature for 30 min then concentratedin vacuo to give 0.076 g of 1-9 as a beige solid.

MS (ESI): m/z=612.3 (MH+).

Examples 2-9

The following compounds were prepared using methods analogous to thosedescribed in the preceding example:

2 R^(a) = H (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-R^(b) = H 17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(2- R^(c) =2-pentyl pentyl)carbamate Mass Spectrum (ESI): m/z = 506.7 (MH⁺) 3 R^(a)= H (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17- R^(b) =H (hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(2-methyl- R^(c) =2-methyl- pentyl)carbamate pentyl Mass Spectrum (ESI): m/z 520.7 (MH⁺) 4R^(a) = H (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-R^(b) = H (hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(3-methyl-2-R^(c) = 3-methyl- propyl)carbamate 2-propyl Mass Spectrum (ESI): m/z =506.7 (MH⁺) 5 R^(a) = COCF₃(11β,16β,17α)-9-fluoro-11-trifluoroacetoxy-16-methyl-3,17a- R^(b) =COCH₃ dioxo-17-[(acety1oxy)methy1]-D-homoandrosta-1,4-dien-17-yl- R^(c)= 1-adamantyl (1-adamantyl)carbamate Mass Spectrum (ESI): m/z =708.3(MH⁺) 6 R^(a) = H(11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17- R^(b) =COCH₃ [(acetyloxy)methyl]-D-homoandrosta-1,4-dien-17-yl-(1- R^(c) =1-adamantyl adamantyl)carbamate Mass Spectrum (ESI): m/z =612.3 (MH⁺) 7R^(a) = COCF₃(11β,16β,17α)-9-fluoro-11-trifluoroacetoxy-16-methyl-3,17a- R^(b) = Hdioxo-17-(hydroxymethyl)-D-homoandrostra-1,4-dien-17-yl-(1- R^(c) =1-adamantyl adamantyl)carbamate Mass Spectrum (ESI): m/z 666.3 (MH⁺) 8R^(a) = H (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo- R^(b)= H 17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl- R^(c) =cyclopentyl (cyclohexyl)carbamate Mass Spectrum (ESI): m/z 504.3 (MH⁺) 9R^(a) = H (11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo- R^(b)= H 17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl- R^(c) = cyclohexyl(cyclohexyl)carbamate Mass Spectrum (ESI): m/z = 518.3 (MH⁺)

Biological Evaluation

The compounds exemplified in the present application exhibited activityin one or more of the following assays.

Ligand Binding Assay Materials:

Binding Buffer: TEGM (10 mM Tris-HCl, 1 mM EDTA, 10% glycerol, 1 mMbeta-mecaptoethanol, 10 mM Sodium Molybdate, pH 7.2)

50% HAP Slurry: Calbiochem Hydroxylapatite, Fast Flow, in 10 mM Tris, pH8.0 and 1 mM EDTA. Wash Buffer: 40 mM Tris, pH7.5, 100 mM KCl, 1 mM EDTAand 1 mM EGTA. 95% EtOH

Dexmethasone-methyl-3H, (DEX*); (Amersham cat# TRK645)Dexamethasone(DEX) (Sigma, cat# D1756):

Hydroxylapatite Fast Flow; Calbiochem Cat#391947 Molybdate=Molybdic Acid(Sigma, M1651) MDA-MB-453 Cell Culture Media:

RPMI 1640 (Gibco 11835-055) w/23.8 mM NaHCO₃, 2 mM L-glutamine in 500 mLof complete media Final conc. 10 mL (1M Hepes) 20 mM 5 mL (200 mM L-glu)4 mM 0.5 mL (10 mg/mL human insulin) 10 μg/mL in 0.01 N HClCalbiochem#407694-S) 50 mL FBS (Sigma F2442) 10% 1 mL (10 mg/mLGentamicin 20 μg/mL Gibco#15710-072)

Cell Passaging

Cells (Hall R. E., et al., European Journal of Cancer, 30A: 484-490(1994)) MDA-MB-453 (ATCC) cultured in RPMI 1640 (Gibco 11835-055)containing 20 mM Hepes, 4 mM L-glu, 10 ug/ml of human insulin (Sigma,I-0259), 10% FBS and 20 ug/ml of Gentamicin (Gibco#15710-072) are rinsedtwice in PBS. Phenol red-free Trypsin-EDTA is diluted in the same PBS1:10. The cell layers are rinsed with 1× Trypsin, extra Trypsin ispoured out, and the cell layers are incubated at 37° C. for ˜2 min. Theflask is tapped and checked for signs of cell detachment. Once the cellsbegin to slide off the flask, the complete media is added. The cells arecounted at this point, then diluted to the appropriate concentration andsplit into flasks or dishes for further culturing (Usually 1:3 to 1:6dilution).

Preparation of MDA-MB-453 Cell Lysate

When the cells are 70 to 85% confluent, they are detached as describedabove, and collected by centrifuging at 1000 g for 10 minutes at 4° C.The cell pellet is washed twice with TEGM (10 mM Tris-HCl, 1 mM EDTA,10% glycerol, 1 mM beta-mercaptoethanol, 10 mM Sodium Molybdate, pH7.2). After the final wash, the cells are resuspended in TEGM at aconcentration of 10⁷ cells/mL. The cell suspension is snap frozen inliquid nitrogen or ethanol/dry ice bath and transferred to −80° C.freezer on dry ice. Before setting up the binding assay, the frozensamples are left on ice-water to just thaw (˜1 hr). Then the samples arecentrifuged at 12,500 g to 20,000 g for 30 min at 4° C. The supernatantis used to set-up assay right away. If using 50 μL of supernatant, thetest compound can be prepared in 50 μL of the TEGM buffer.

Procedure for Multiple Compound Screening

1×TEGM buffer is prepared, and the isotope-containing assay mixture isprepared in the following order: EtOH (2% final concentration inreaction), ³H-DEX (Amersham Biosciences) and 1×TEGM. [e.g. For 100samples, 200 μL (100×2) of EtOH+4.25 μL of 1:10 ³H-Dex stock+2300 μL(100×23) 1×TEGM]. The compound is serially diluted, e.g., if startingfinal conc. is 1 μM, and the compound is in 25 μL of solution, forduplicate samples, 75 μL of 4×1 μM solution is made and 3 μL of 100 μMis added to 72 μL of buffer, and 1:5 serial dilution.

25 μL of ³H-DEX (6 nM) trace and 25 μL compound solution are first mixedtogether, followed by addition of 50 μL receptor solution. The reactionis gently mixed, spun briefly at about 200 rpm and incubated at 4° C.overnight. 100 μL of 50% HAP slurry is prepared and added to theincubated reaction which is then vortexed and incubated on ice for 5 to10 minutes. The reaction mixture is vortexed twice more to resuspend HAPwhile incubating reaction. The samples in 96-well format are then washedin wash buffer using The FilterMate™ Universal Harvester plate washer(Packard). The washing process transfers HAP pellet containingligand-bound expressed receptor to Unifilter-96 GF/B filter plate(Packard). The HAP pellet on the filter plate is incubated with 50 μL ofMICROSCINT (Packard) scintillint for 30 minutes before being counted onthe TopCount microscintillation counter (Packard). IC₅₀s are calculatedusing DEX as a reference.

Examples 1 to 9 were tested in the ligand binding assay and demonstratedIC₅₀s less than 1000 nM.

Trans-Activation Modulation of Glucocorticoid Receptor (GRAMMER)

This assay assesses the ability of test compounds to controltranscription from the MMTV-LUC reporter gene in lung adenocarcinomaA549 cells or MDA-MB-453 cells, a human breast cancer cell line thatnaturally expresses the human GR. The assay measures induction of amodified MMTV LTR/promoter linked to the LUC reporter gene.

The routine transient assay consists of plating 7,000-25,000 cells/wellof a white, clear-bottom 96-well plate. Alternatively, 384-well platescan be used at a cell concentration of 10,000/well. The media that thecells are plated in is “exponential growth medium” which consists ofphenol red-free RPMI1640 containing 10% FBS, 4 mM L-glutamine, 20 mMHEPES, 10 ug/mL human insulin, and 20 ug/mL gentamicin. Incubatorconditions are 37° C. and 5% CO₂. The transfection is done in batchmode. The cells are trypsinized and counted to the right cell number inthe proper amount of fresh media. It is then gently mixed with theFuGene6/DNA mix and plated onto the 96 or 384-well plate, all the wellsreceive 100 uL or 40 uL, respectively, of medium+lipid/DNA complex thenincubated 37° C. overnight. The transfection cocktail consists ofserum-free OptiMEM, FuGene6 reagent and DNA. The manufacturer's (RocheBiochemical) protocol for cocktail setup is as follows: The lipid to DNAratio is approximately 2.5:1 and the incubation time is 20 min at roomtemperature. Sixteen to 24 hours after transfection, the cells aretreated with dexamethasone to a final concentration of 10 nM as well asthe compound of interest, such that final DMSO (vehicle) concentrationis equal to or less than 1%. Each plate also contains samples that aretreated with 10 nM dexamethasone alone, which is used as the 100%activity control. The cells are exposed to the compounds for 24 hours.After 24 hours, the cells are lysed by a Promega cell culture lysisbuffer for approximately 30 min and then the luciferase activity in theextracts is assayed in the 96-well format luminometer. In 384-wellformat, Steady-Glo (Promega) or Steady-Lite (PerkinElmer) can be used byadding an equal volume of reagent to the media present in each well.Activity induced by 10 nM dexamethasone alone is set at 100% activity.Antagonist activity is calculated by determining the decrease indexamethasone-induced activity in response to compound treatmentrelative to samples that were treated with dexamethasone alone. Resultsare expressed as % inhibition of 10 nM dexamethasone activity or as foldof 10 nM dexamethasone activity. This transactivation assay can beperformed in an agonist and antagonist mode to identify these differentactivities.

Activity of test compounds is calculated as the E_(max) relative to theactivity obtained with 300 nM dexamethasone. Activity of test compoundsis calculated as the E_(max) relative to the activity obtained with 300nM DEX. The exemplified tissue selective glucocorticoid receptormodulators of the present invention display partial agonist activity inthis assay of greater than 5%.

The action of compounds is also tested in an antagonist mode(Anti-GRAMMER) in which the cells are treated with medium containing anagonist such as 10 nM DEX and the ability to agents to inhibit theactivation by an agonist is measured.

Transrepression Assay

This assay assesses the ability of test compounds to controltranscription from the TNFα-β-lactamase reporter gene in U937 cells, ahuman myelomonocytic leukemia cell line that naturally expresses thehuman GR. The assay measures compound dependent-repression of the TNFαpromoter linked to a reporter gene.

The human U937 cells that had been stablely transfected with theTNF-αpromoter driving β-lactamase are used for this assay. U937 cellscontain an endogenous glucocorticoid receptor (GR). Cells are maintainedin RPMI 1640 Growth medium (Gibco Cat#11875-093) containing 25 mM HEPES,10% FBS, 2 mM L-Glutamine, 1 mM Sodium pyruvate, 25 μg/ml Gentamicin(Gibco Cat#15710-064), 1:1000 2-Mercaptoethanol (Gibco Cat#21985-023)and 0.8 mg/ml G418 (Gibco Cat#10131-027). The density of the cells inthe flask needs to be about 1×10⁶−3×10⁶/ml at the time of harvest.Usually, the cells are split to 1.2˜1.4×10⁵/ml (1:10) 3 days prior tothe assay. 50,000 cells/well are plated in 96 well black-walled platesthe day of assay. Test compounds are added 10 μL/well, and cells areincubated at 37° C. for 30˜45 min. For assaying compounds, first dilute1:10 in DMSO to make 1 mM, then further dilute 1:100 in medium to make10× stock prior to adding to the cells. Add 50 ng/ml PMA (Sigma, cat#P8139) 10 μL/well to a final concentration 5 ng/ml, and 1 μg/ml LPS(Sigma, cat# L4130) 10 μL/well to a final concentration 100 ng/ml.Incubate cells at 37° C. overnight for ˜18 hr. PMA is stored frozen as100 μg/ml stock in DMSO. Dilute 1:10 in DMSO for a working stock of 10μg/ml and store at −20 C. For assaying, dilute the 10 μg/ml workingstock 1:200 in medium to make a 10× solution (50 ng/ml). Store frozenLPS at 1 mg/ml in PBS, dilute 1:1000 in medium to make 10× (1 μg/ml) forthe assay. Add 6× loading buffer (CCF₂-AM) 20 μL/well, and incubate atroom temperature for 70˜90 min. Read plates on CytoFluor II Plate Readeraccording to manufacture suggested protocols. The activity repressed by100 nM dexamethasone alone is set as 100% activity.

Examples 1 to 9 were tested in the transrepression assay anddemonstrated maximal activity greater than 5%.

Microarray Analysis

All cell culture reagents were purchased from Invitrogen Life Tech,Carlsbad Calif. A549 cells were grown in phenol red-free DMEM/F12 mediumsupplemented with 10% FBS. Cells were grown at 37° C. with 5% CO₂. Usingthe RNeasy Kit (Qiagen Corp, Valencia Calif.), total RNA was extractedand purified from A549 cells treated with different GC compounds for 24hours, at a fully active dose. These cells express large amount of theGR and are very responsive to GC treatment. All samples were comparedagainst cells treated with vehicle. Expression levels of 23000 geneswere measured using oligonucleotide microarrays purchased from AgilentTechnologies, Inc. Each comparison was done on a pair of microarrayswith reversed fluorophores. Raw image intensity data were processedaccording to the method described in U.S. Pat. No. 6,351,712. The methodwas used to remove dye bias and to derive a Rosetta probability (p) andfold change value for each gene and each sample pair. Furthermore, foreach gene an ANOVA model was constructed across all treatments to deriveerror estimates. P values for evaluating expression differences werecomputed using a Bayesian adjusted t-test that was developed byLönnstedt and Speed (2002) and extended by Smyth (2003). A gene wasdeclared differentially expressed in any particular comparison if itsatisfied two critera:

1. The Rosetta p value had to be less than 0.1 and the Rosetta foldchange value had to be greater than 1.4 in at least one of thetreatments.

2. The ANOVA p value had to be less than 0.01 and the fold changegreater than 2 in the comparison under consideration.

In Vivo Inflammation Assay

Intact adult (6 month old) female Sprague-Dawley rats are used in theoxazolone (OX) contactdermatitis model. Rats were sensitized on theventral abdomen with OX on Day 0. On Days 7 and 9, a randomly-selectedear was challenged (same ear each time) with OX; the other was treatedwith vehicle. Daily treatment begun on Day 7 and continued for 7d withtest compounds at different doses and 1.3 mpk 6-methlyprednisolone or0.1 mpk DEX as positive controls. The thickness of both ears aremeasured on Days 11 and 14. Necropsy occurred on Day 14. The rat isfirst weighed, then anesthetized in a CO₂ chamber until near death.Approximately 5 ml whole blood is obtained by cardiac puncture. The ratis then examined for certain signs of death and completeness. Tissuesare dissected in a highly stylized fashion. The following endpoints wereevaluated: a) inhibiting ear inflammation induced by oxazalone, b)raising serum insulin, c) reducing serum ACTH, d) reducing spleenweight, e) reducing skin thickness, f) reducing body weight, g)increasing expression of bone-related genes with potential relationshipto negative glucocorticoid effects on bone; e) changes in molecularmarkers that correlate with skin inflammation, skin thinning, muscleatrophy and glucose metabolism in liver. All blood samples werecollected between 1330-1530 hours, ˜4-5 hrs after the last compoundtreatment.

Primary data for this assay are left and right ear thickness. Inter-earthickness difference (etd) is used for the estimating the level ofinflammation and effectiveness of the compounds is determined by theirability to reduce the increase the thickness of the inflamed ear. Backof the rat skin thickness, spleen weight, serum insulin as well as theeffects of gcs on the expression of molecular markers in skininflammation, skin atrophy, muscle atrophy and glucose metabolism inliver are measured. Data are analyzed by anova plus fisher plsd post-hoctest to identify intergroup differences.

1. A compound of Formula I

or a pharmaceutically acceptable salt thereof, wherein R¹ and R² areindependently selected from the group consisting of: R^(a), —C(O)—R^(a),and —C(O)—N(R^(a))R^(a); R³ and R⁴ are independently selected from thegroup consisting of: hydrogen, C₁₋₁₀alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,C₃₋₁₀cycloalkyl, aryl, hetreoaryl, heterocyclyl, and C₁₋₁₀fluoroalkyl;and each Ra is independently selected from the group consisting of:hydrogen, C₁₋₁₀alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₃₋₆cycloalkyl, aryl,hetreoaryl, heterocyclyl, and C₁₋₁₀fluoroalkyl.
 2. The compoundaccording to claim 1 wherein R² is selected from hydrogen and CH₃—C(O)—.3. The compound according to claim 1 wherein R¹ is selected fromhydrogen and CF₃—(CO)—.
 4. The compound according to claim 1 wherein R³is hydrogen.
 5. The compound according to claim 4 wherein R⁴ is selectedfrom C₁₋₁₀alkyl and C₃₋₁₀cycloalkyl.
 6. The compound according to claim1 wherein: R¹ is selected from hydrogen or CF₃—(CO)—; R² is selectedfrom hydrogen and CH₃—C(O)—; R³ is hydrogen; and R⁴ is selected fromC₁₋₁₀alkyl and C₃₋₁₀cycloalkyl.
 7. The compound according to claim 6wherein R⁴ is C₁₋₆alkyl.
 8. The compound according to claim 6 wherein R⁴is selected from: cyclopentyl, cyclohexyl and adamantanyl.
 9. Thecompound according to claim 6 wherein R¹ and R² are hydrogen.
 10. Thecompound according to claim 1 selected from the following group:(11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate,(11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(2-pentyl)carbamate,(11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(2-methyl-pentyl)carbamate,(11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(3-methyl-2-propyl)carbamate,(11β,16β,17α)-9-fluoro-11-trifluoroacetoxy-16-methyl-3,17a-dioxo-17-[(acetyloxy)methyl]-D-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate,(11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-[(acetyloxy)methyl]-D-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate,(11β,16β,17α)-9-fluoro-11-trifluoroacetoxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(1-adamantyl)carbamate,(11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(cyclopentyl)carbamate,(11β,16β,17α)-9-fluoro-11-hydroxy-16-methyl-3,17a-dioxo-17-(hydroxymethyl)-D-homoandrosta-1,4-dien-17-yl-(cyclohexyl)carbamateor a pharmaceutically acceptable salt of any of the above.
 11. Apharmaceutical composition comprising a compound according to claim 1 incombination with a pharmaceutically acceptable carrier.
 12. A method fortreating a glucocorticoid receptor mediated disease or condition in amammalian patient in need of such treatment comprising administering thepatient a compound according to claim 1 in an amount that is effectivefor treating the glucocorticoid receptor mediated disease or condition.13. The method according to claim 12 wherein the glucocorticoid receptormediated disease or condition is selected from the group consisting of:tissue rejection, leukemias, lymphomas, Cushing's syndrome, acuteadrenal insufficiency, congenital adrenal hyperplasia, rheumatic fever,polyarteritis nodosa, granulomatous polyarteritis, inhibition of myeloidcell lines, immune proliferation/apoptosis, HPA axis suppression andregulation, hypercortisolemia, stroke and spinal cord injury,hypercalcemia, hypergylcemia, acute adrenal insufficiency, chronicprimary adrenal insufficiency, secondary adrenal insufficiency,congenital adrenal hyperplasia, cerebral edema, thrombocytopenia,Little's syndrome, obesity, metabolic syndrome, inflammatory boweldisease, systemic lupus erythematosus, polyartitis nodosa, Wegener'sgranulomatosis, giant cell arteritis, rheumatoid arthritis, juvenilerheumatoid arthritis, uveitis, hay fever, allergic rhinitis, urticaria,angioneurotic edema, chronic obstructive pulmonary disease, asthma,tendonitis, bursitis, Crohn's disease, ulcerative colitis, autoimmunechronic active hepatitis, organ transplantation, hepatitis, cirrhosis,inflammatory scalp alopecia, panniculitis, psoriasis, discoid lupuserythematosus, inflamed cysts, atopic dermatitis, pyoderma gangrenosum,pemphigus vulgaris, buflous pemphigoid, systemic lupus erythematosus,dermatomyositis, herpes gestationis, eosinophilic fasciitis, relapsingpolychondritis, inflammatory vasculitis, sarcoidosis, Sweet's disease,type I reactive leprosy, capillary hemangiomas, contact dermatitis,atopic dermatitis, lichen planus, exfoliative dermatitus, erythemanodosum, acne, hirsutism, toxic epidermal necrolysis, erythemamultiform, cutaneous T-cell lymphoma, Human Immunodeficiency Virus(HIV), cell apoptosis, cancer, Kaposi's sarcoma, retinitis pigmentosa,cognitive performance, memory and learning enhancement, depression,addiction, mood disorders, chronic fatigue syndrome, schizophrenia,sleep disorders, and anxiety.
 14. The method according to claim 12wherein the glucocorticoid receptor mediated disease or condition isselected from the group consisting of: tissue rejection, Cushing'ssyndrome, inflammatory bowel disease, systemic lupus erythematosus,rheumatoid arthritis, juvenile rheumatoid arthritis, hay fever, allergicrhinitis, asthma, organ transplantation, inflammatory scalp alopecia,psoriasis, discoid lupus erythematosus, and depression.
 15. (canceled)16. (canceled)